权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Development of a New Thermostable Photosynthetic Bactria by Ricombinant DNA

利用重组DNA开发新型耐热光合细菌

基本信息

批准号：
63044016
负责人：
NOZAWA Tsunenori
金额：
$ 2.37万
依托单位：
TOHOKU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Overseas Scientific Survey.
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1989
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-63044016/
关键词：
Photosynthetic bacteria Thermostability Recombinant DNA Primary process of photosynthesis Reaction center protein Base sequence of DNA Crystallization of protein X-ray crystal analysis of protein

项目摘要

In order to develop a new thermostable photosynthetic bacterium, we have investigated the origin of the thermostability in the photosynthetic reaction, and cloning of the photosynthetic reaction center gene for point mutation by recombinat DNA technique.Comparison of protein structures between thermostable photosynthetic bacteria (Chloroflevus aurantiacus, Chromatium tepidum), and mesophilic ones (Chromatium vinosm, Rhodobacter sphaeroides) gave a key to understand the origin. Thermostabel reaction center protein were revealed to have extra structures to keep their thrtiary structure rigid.Genomic DNA was extracted from whole cell of a thermophilic purple sulfur bacterium Chromatium tepidum. From genomic DNA digested by a restriction enzyme Sau 3Al, a gene bank (consist of 800 colonies) was produced by in vitro packaging and transfection to a Ecoli cell. A clone which encodes the photosynthetic reaction center protein for the primary reaction of the photosynthesis was selected and under analysis of the DNA base sequence.Purification of the reaction center protein and trial for crystallization was carried out in order to get three dimensinal structures which are needed to pin point the place to make point mutation for improving thermal stability.

为了培育新的耐热光合细菌，我们研究了光合反应耐热性的来源，并利用重组DNA技术克隆了光合反应中心基因进行点突变。比较耐高温光合细菌（Chloroflevus aurantiacus, Chromatium tepidum）和中温光合细菌（Chromatium vinosm, Rhodobacter sphaeroides）的蛋白质结构为了解其起源提供了关键。热稳定反应中心蛋白具有额外的结构以保持其三级结构的刚性。从嗜热紫硫菌（Chromatium tepidum）的全细胞中提取基因组DNA。用限制性内切酶Sau 3Al消化基因组DNA，通过体外包装和转染到Ecoli细胞中，产生了一个由800个菌落组成的基因库。选择了一个编码光合作用初级反应中心蛋白的克隆，并对其DNA碱基序列进行了分析。对反应中心蛋白进行了纯化和结晶试验，获得了为提高热稳定性而定点突变所需的三维结构。

项目成果

期刊论文数量（14）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Tsunenori Nozawa, Mamoru Terauchi, Takayoshi Kobayashi, Masahiro Hatano: "Picosecond Absorption Spectra of a Reaction Center from a Novel Thermophilic Photosynthetic Bacterium Chromatium tepidum" Springer Series in Chemical Physics, Ultrafast Phenomena VI

Tsunenori Nozawa、Mamoru Terauchi、Takayoshi Kobayashi、Masahiro Hatano：“新型嗜热光合细菌 Chromatium tepidum 反应中心的皮秒吸收光谱”施普林格化学物理系列，超快现象 VI

DOI：
发表时间：
期刊：
影响因子：
0
作者：
通讯作者：

Masaru Ohta, Tsunenori Nozawa, Masahiro Hatano, Hidenori Hayashi, Mitsuo Tasumi, Keizo Shimada: "^<31>P-NMR studies of photophosphorylation in Chromatophore from Chromatium vinosum" Biochimica et Biophysica Acta. 1011. 33-39 (1989)

Masaru Ohta、Tsunenori Nozawa、Masahiro Hatano、Hidenori Hayashi、Mitsuo Tasumi、Keizo Shimada：“来自 Chromatium vinosum 的色素细胞中光磷酸化的^ 31 P-NMR 研究”Biochimica et Biophysicala Acta。

DOI：
发表时间：
期刊：
影响因子：
0
作者：
通讯作者：

Tsunenori Nozawa, Hideki Tanaka, Masayuki Kobayashi, Joe W. Farcus, Friedrich Lottspeich, Dieter Oesterhelt, Charles Eberhart: "Characterization of amino acid sequence of a photosynthetic bacterium Chromatium tepidum and origin of the thermostability dedu

Tsunenori Nozawa、Hideki Tanaka、Masayuki Kobayashi、Joe W. Farcus、Friedrich Lottspeich、Dieter Oesterhelt、Charles Eberhart：“光合细菌 Chromatium tepidum 氨基酸序列的表征以及热稳定性推论的起源

DOI：
发表时间：
期刊：
影响因子：
0
作者：
通讯作者：

Tsunenori Nozawa,Jeffrey T.Trost,Taisei Fukada,Masahiro Hatano,James D.McManus,Robert E.Blankenship: "Properties of the reaction center of the thermophilic purple photosynthetic bacterium as Chromatium tepidum" Biochimica et Biophysica Acta. 894. 468-476

Tsunenori Nozawa，Jeffrey T.Trost，Taisei Fukada，Masahiro Hatano，James D.McManus，Robert E.Blankenship：“嗜热紫色光合细菌作为 Chromatium tepidum 的反应中心的特性”生物化学和生物物理学学报。